Anti microbial compositions containing unsymmetrical oligoquaternary ammonium compounds

ABSTRACT

Unsymmetrical oligoquaternary ammonium compounds containing at least two onium nitrogen atoms. Typical compounds are: (C12H25N (Me)2(CH2)4N (Me)2CH2C6H5)Br; (C14H29N (Me)2(CH2)10N (Me)2CH2C6H4Cl)Br2 AND (C12H25N (Me)2(CH2)4N (Me)2(CH2)2N (Me)2CH2C6H5)Br3 These compounds are effective as antimicrobial agents and as anticaries agents.

United States Patent Bauman Dec. 9, 1975 ANTI MlCROBlAL COMPOSITIONSCONTAINING UNSYMMETRICAL OLIGOQUATERNARY AMMONIUM COMPOUNDS RobertAndrew Bauman, New Brunswick, NJ.

Colgate-Palmolive Company, New York, N.Y.

Filed: Sept. 17, 1974 Appl. No.: 506,807

Related US. Application Data Continuation of Ser. No. 185,388, Sept. 30,1971. abandoned, which is a division of Ser. No. 82,594, Oct. 21, 1970,abandoned, which is a continuation-in-part of Ser. No. 716,412. March27.

1968, abandoned.

lnventor:

Assignee:

US. Cl. 424/329; 260/5676; 424/54 Int. Cl. A61K 7/22; A61K 31/14 Fieldof Search 424/54, 329

References Cited UNITED STATES PATENTS 4/1938 Taub et aliiiiiiiiiiiiiiiiiiiiii 260/5676 3,489,663 l/1970 Bayer et a1 204/59OTHER PUBLICATIONS J. Von Braun et al., Ber. 49: 2610-2612 (1916); J.Chem. Soc. 1121175 (1917).

A. P. Gray et al., Nature 181: 634-635 (1958).

F. Vidal, J. Org. Chem. 24: 680-683 (1959).

J. L. Hartwell et al., J. Am. Chem. Soc. 72: 2040-2044 (1950).

T. P. Abbiss et al., J. Chem. Soc. 2248-2254 (1964).

Primary ExaminerShep K. Rose Attorney, Agent, or Firm Robert L. Stone;Herbert S. Sylvester; Murray M. Grill [57] ABSTRACT 14 Claims, NoDrawings t 1 R2 R2 This application is a continuation of applicationSer. No. 185,388, filed Sept. 30, 1971, which was a division of 9 medwh'ch was 3 wherein R is a substantially straight chain alkyl groupcommuauonqmlgan, filed having to 18 carbon atoms; Y is selected from theI968, all three applications being now abandoned. group Consisting ofm+p)] RmsXp and The present invention relates to unsymmetrical ohgo- 10quaternary ammonium compounds and, more particu larly, to unsymmetricaloligoquaternary ammonium ((142mm+ CH2CH[Fmwm]RmtixpL compoundscontaining at least two omumnitrogen atoms, having anti-microbialactivity, having anti-caries R" propertiesand having novel adsorptionand desorption 5 properties. it is an integer 2 to 18; R, R R and R arelower al- In recent years a multitude of quaternary ammonium kyls having1 to 3 carbon atoms; b is an integer 2 to 12; compounds havingantifungal and antibacterial capa- X is selected from the groupconsisting of chlorine, brobilities have been disclosed. Thesequaternary ammomine and iodine; m is O to 3; p is O to 3; m+p is 0 to3', nium compounds may be divided into two major groups 20 An is acompatible anion, including halide, such as Cl, (A) those quatemarieshaving only one onium nitrogen Br, or alkyl or phenyl sulfonate, such asCH SO atom and (B) those quatemaries having more than one C H SO p-CH CH SO etc. in sufficient number onium nitrogen atom. The latt r gr ca b sbdito satisfy the positive valence of the remainder of the vided intobisquaternaries having two onium nitrogen compound, that is, 2 or 3. TheAn groups may be the atoms and tri and higher quatemaries having threeor same or different. more onium nitrogen atoms. Each of thesesubdivisions Illustrative of the effect of the value of n in the forofthe poly-onium nitrogen atom quatemaries can be mula set forthhereinbefore are the following data obfurther divided into thesymmetrical and the unsymtained in anti-microbial tests employing thefollowing metrical poly-onium nitrogen atom quatemaries deorganisms:pendent upon the substituent groups attached to the terminal oniumnitrogen atoms of the poly-onium nitroorganism gen atom quatemaries.Thus, when all of the substituent groups attached to each terminal oniumnitrogen g f 'i i f f i urynehacrerium acne: (gram positive bacteria)atom are the same, the quaternary is considered to be 3 Ban-11m sum/ii(gram positive bacteria) symmetrical. When one or more of thesubstituent 4 ESC'w'iv/Iifl wli a negative baclefial groups attached toone terminal onium nitrogen atom Z jfjjjfgj'ggj jffjfijggf F banana) ofa poly-onium nitrogen atom quaternary is or are dif- 7 Tri wphymn mmagmphyw (f ng ferent from the constituent groups attached to the g gfif "i ii pmmccu: mm: (5-3) (gram positive other terminal onium nitrogenatom the quaternary is bacteria) considered to be unsymmetrical orasymmetrical. 4O

Thqsgskmed m the recogmze that 9 an The serial dilution test todetermine the minimal indescriptions of poly-onium compounds do notindicate hibitory concentration in Lg/ml Le. l x mg/ml, that the emcacydependent q the .number of has been used to illustrate this importantcharacteristic groups which separate the two onium-nitrogen atoms in ofthese novel poly onium quaternary ammonium type the bls'quafemanes thatthe poly-0mm commaterials. Two materials of this type, A and B, werepounds having three onium-nitrogen atoms that the efused and the chainlength (n) varied from 3 to 10 ficacy is dependent upon the number of CHgroups (CH 2 groups. separating from the central onium-nitrogen atom theterminal onium-nitrogen atom to which is bonded an alkyl group havingten or more carbon atoms. It has A now been discovered that the numberof CH -groups Me MeM Me separating the aforesaid onium-nitrogen atoms isindic- H C H ative of the efficacy of the germicidal properties and a I2 a 5 other properties such as cariostatic properties of the Compound Bquaternary compound. In addition, the substantivity of theseunsymmetrical quatemaries decreases as the con- Me Me Me /Me Me /Mecentration of methanol decreases in aqueous acid contaming methanol inproportions varying from O to 55 Br, percent as will be discussed belowwith regard to chromatography tests.

The novel poly-onium nitrogen compounds of the In the above compounds, Aand B as well as in strucpresent invention can be represented by theormu tural formulae set forth hereinafter, Me indicates a methyl group.

Compound A Organism No. l 2 3 4 5 r 7 s n p g/ml pig/ml pg/ml Lg/mlug/ml ug/ml pig/ml ngiml Compound A-continued Organism No. I 2 3 4 5 6 78 3 25 2.5 25 100 100 12.5 125 1000 4 50 100 12.5 100 100 25 10011 50116 12.5 0.78 6.25 25 250 1.56 71; 25() 8 1.56 0.2 6.25 3.12 10011 1.567.8 312 10 0.78 (1.1125 1.56 12.5 100 0.78 7.8 31.3 Compound B Compoundshaving values of n, ranging from low to high, are effectiveantimicrobial agents. Generally, the compounds of greater effectivenesshave higher values of n. Similar results may be observed when the methylgroups are replaced with ethyl and/or propyl groups.

The effect of a single nuclear substituent, such as a halogensubstituent in the benzyl ring is clearly shown by the followingcomparative results using the serial dilution test and reporting theminimal inhibitory concentration in ag/ml, that is, 1X10 mg/ml oncompounds in which the chain length (n) was 4:

alkyl group such as n-tridecyl and longer alkyl groups such asn-pentadecyl, n-hexadecyl, n-heptadecyl and n-octadecyl.

In addition to the antimicrobial effectiveness of compounds of theinvention, compounds of the invention are also effective in reducingformation of caries in mammals as demonstrated below:

Caries-susceptible hamsters bred either from the Keyes strain or theNlDR (National lnstitute for Dental Research) strain in groups of 15males and 15 females per control group and per each test group were fedad A nC -,,H NMe,(CH N*Me,CH C,,H,pC1

Bi Organism No. l 2 3 4 5 6 7 8 p-glml pig/ml [Lg/ml u-g/ml pig/m1pig/ml g/ml ig/ml A 50 100 12.5 100 100 1000 500 Alml 6.25 0.78 6.25 251110 12.5 31.3 125 Compound 8 v. Compound B,,,,, B nC H N*Me,(CH,NMe,(CH,),NMe CH,C H,

B nC,,H- N*Me ,(CH,).N*Me,(CH,),N*Me,CH,C,,H,p-Cl

Br,Cl Organism #g/ml pig/m1 pig/m1 pig/ml pg/ml pg/ml pig/m1 pg/ml B3.13 3.13 100 25 1000 B 12.5 6.25 0.78 25 100 25 15.6 2511 Generallyspeaking the presence of a nuclear substituent such as a halogensubstituent in the benzyl ring increases the antimicrobial efiectivenessof compounds of the invention. In addition to or in place of the halogensubstituent, alkyl substituent, such as methyl, ethyl or propyl, may bepresent in the benzyl ring.

The following compounds were also tested against the organisms describedabove and the inhibitory concentration indicated results below wereobtained:

Compound C lib a Mitchell cariogenic diet and received constantdeionized water. Each day each hamsters teeth were swabbed 30 seconds bycotton tipped swabs, the control group with water and each test groupwith its test solution. After 6 weeks of swabbing the animals weresacrificed, and the defleshed heads were scored by a modified version ofthe Keyes scoring method. Mean averages and percentage changes from thecontrol group were determined and tested statistically to deter- No. l 23 4 5 6 7 8 9 pg/ml pig/ml ug/ml ug/ml pig/m1 ag/rnl pig/ml pig/m1pig/m1 Chnl 0.78 3.12 0.39 1.56 62.5 (1.78 31.2 250 3.12 D 0.39 3.120.19 0.78 31.2 11.7% 1.9 62.5 (1.39

Similar results may be obtained when the dodecyl and tetradecyl groupsof Compounds C and D are replaced by shorter substantially straightchain alkyl groups such as n-decyl or n-undecyl, an intermediate minethe significance.

The compounds were administered in 1% by weight solutions.

Results Hamster Caries Test Caries Reduction '7:

The results above indicate the significant effectiveness of thecompounds of the invention in reducing caries formation.

When used against bacteria and fungi, compounds of taining ion such assodium fluoride, stannous fluoride and sodium monofluorophosphate.

The oral preparation may also be a liquid such as a mouth rinse whichtypically contains 20-99% by weight the instant invention may be applieddirectly to the sur- 5 of an aqueous alcohol vehicle, the alcohol beinga cosface to be protected or may be used in a pharmaceutimeticallyacceptable and nontoxic alcohol such as ethcal carrier. Typically aneffective amount, e.g. 0.1 to anol or isopropyl alcohol and beingpresent in amount about l% by weight of the quaternary is included in anof about -30% by weight of the oral preparation. inert liquid carriersuch as water and a dispersing or Such oral preparations are typicallyapplied by brushsurface active agent. Alternatively, an effective ingthe teeth or rinsing the oral cavity for -90 secamount, e.g. O.l toabout l0% by weight may be incoronds at least once daily. Typical oralpreparations inporated into a solid carrier which may be inert, such ascluding a compound of the instant invention are set talc, clay,diatomaceous earth, flour, etc. forth as follows:

Dental Cream Mouth Wash Parts Parts Component By Weight Component ByWeight Quaternary D,,,,, 0.5 Quaternary C,,,,, 0.25

Sodium Benzoate 0.15 Ethyl Alcohol H.782

Saccharine 0.2 Flavor 0.3 l 8 insoluble Sodium 42.] Glycerine l0.00(l

Metaphosphate Dicalcium Phosphate 5.0 Deionized Water 74.]

Dihydrate Titanium Dioxide 0.4 Color 0.55

(l% Solution) Gurn Tragacanth l.4

Oil of Wintergreen l.0

Color 0.03

Water 22. i2

Glycerine 99.3%) 27. l0

When compounds of the invention are used in compositions which reduceformation of caries they are typically incorporated in oral preparationsin effective amounts up to about 5% by weight, preferably 0.1% and mostpreferably 0.25-0.5% by weight of the oral preparation. Typically theoral preparation is a dentifrice such as a dental cream, tablet orpowder, containing as a vehicle about 20-95% by weight of a dentallyacceptable water-insoluble polishing material, preferably includingwater-insoluble phosphates such as dicalcium phosphate, tricalciumphosphate, insoluble sodium metaphosphate and calcium pyrophosphate. Thedentifrice may also include water, binders such as glycerine, sorbitol,propylene glycol and polyethylene glycol 400, gelling agents such asIrish moss and sodium carboxymethyl cellulose, additional antibacterialagents, coloring or whitening agents, preservatives, silicones,chlorophyll compounds, additional ammoniated materials, flavoring orsweetening materials and dentally beneficial compounds which providefluorine-con- 6 Br Br The novel poly-onium quaternary ammonium compoundsof the present invention can be prepared by the methods of organicchemistry known by those skilled in the art. Presently, it is preferredto obtain these novel poly-onium compounds by first preparing aquaternary ammonium compound by the reaction of a tertiary amine with analkylene dihalide under suitable reaction conditions. The reaction canbe illustrated by the following equation:

The trialkyl (w-haloalkyl) ammonium bromide is then reacted with (l)dialkyl benzyl amine or (2) dialkyl halobenzylamine or (3) alkylateddialkylbenzylamine or (4) dialkyl polyhalobenzylamine as can berepresented by the following equation:

7 8 (The symbols R, R, R R, X, n, m, and p in the foreb is 2 or 3, n is3 to ID and is greater than b, m is to going equations have the samesignificance as hereinv P is 0 t0 +p l8 0 and Hill is 8 l de hile beforeinitially defined.)

The unsymmetrical or asymmetrical bisquaternary amines in which R is n-CH n is 3, 4, 6, 8, and 10; m is 0, p is l and X is chlorine, have beenprepared in ac- R, R, R R, R and X have the same significance ashereinbefore are prepared from the backbone, trialkylhaloalkylhalide,

cordance with the method set forth in Example 1. R\ R EXAMPLE 1 1 rn-dodecyldimethylamine was reacted with a large ex- Hal Hal cess ofBr(CH. ),,Br in water at 40C. to formndodecyldimethyl-(omega-bromoalkyl) ammonium broid Thi quat ammo i b idewa re- That is to say the trialkylhaloalkylhalide is reacted with fluxedin water with a twofold excess of dimethyl-p- N,N.N',N'tetraalkylalkylene diamine, e.g., N,N,N,- chlorobenzylamine. The products, thebisquaternary N'-tetramethylethylenediamine to produce poly-oniumammonium bromides in which n is 3, 4, 6 and 8, were halide as indicatedby the following equation to obtain R R R-' R R R R R R R R R' x RN*-((H,),,Hal +N-(CH- -JI,N- R-N((H2|,,N-(CH2|,,-N

Hal Hal Hal recrystallized from acetone. The unsymmetrical or 25 Thepoly-onium halide is then reacted with a benzyl asymmetricalbis-quaternaries wherein R is n-C Hzrg halide to produce the poly-oniumquaternary com- R is CH R is CH n is 3, 4, 6, 8 and l0; m is 0', andpound represented by the formula t 2 RI t p is 0 were observed to havethe following characteriswhere R, R, R R, X and Hal, n, b, m, and p havethe tics closely akin to the calculated characteristics: samesignificance as hereinbefore indicated in this paragraph. Illustrativeof the preparation of these latter polygr Cal. N 5.09% Cal. oniumquaternary ammonium compounds is the pron=4 v N 50017 duction of thequaternaries represented by the formula Br 28.32% Cal. N 4.96% Cal. Br23.00% Obs. N 4.84% Obs, n=6

Br 26.97% Cal. Me Me Me Me Me Me Br-27.l7% Obs. r-r 2s lCH- ),,N'(CH I-N'-CH Arr,

Br 25.75% Cal. Br 25,16'7r Obs. n=l0 Br 2464a Cal. N 4.32% Cal. Br 423.68% Obs. N 3.96% Obs.

where n is 4 and l() and An has the same significance as hereinbeforc;and the quaternary represented by the These compounds are furthercharacterized by the formula chromatographic properties hereinbeforedescribed.

Compounds including l-p-chlorophenyl-2,2,l3,l3- Me Me Me MEML. MCtetra-methyl-2,l3-diazonia heptacosane dibromide [n C CH CH u A C H N(Me) (CH N (Me) CH C H p-Cl] Br We F have been similarly prepared usingtetradecyldimethyl ifii f g3821;233:2322; g iggi gg a m s sgl wherein Anhas the same significance as hereinbefore.

l 2. dibromide [Br(CH ),,Br] and the reaction product re- Thesepreparations are set mrth m Exdmp c acted with tetradecyldimethyl amine.EXAMPLE 2 The polyonium quaternary ammonium compounds (:0

represented y the formula n-Dodccyldimethyl-omega-bromo-alkyl halide wasreacted with N,N,N',N'-tetramethylethylenediamine at 25C. in the molalratio of 1:8. The resulting bis-quater- R R R R nary was recrystallizedfrom acetone-chloroform. The

[ H recrystallized bis-quaternary was reacted separately ACH'JL'TN iwith benzyl bromide and chloro-benzyl bromide at 25C. to obtain thepoly-onium quaternary ammonium where Y is 1CH N' -LH bromide Thepoly-onium quaternary ammonium bro- P mides thus prepared have thefollowing characteristics:

Cal Obs.

Bromine 29.94% 28.6371 Nitrogen 5.25% 4.93%

Cl] Br Cl Cal. Obs.

Total Halogen Although this invention has been described with referenceto specific examples, it will be apparent to one skilled in the art thatvarious modifications may be made thereto which fall within its scope.

What is claimed is:

1. An antimicrobial composition comprising about (ll-% by weight of anunsymmetrical poly-onium quaternary ammonium compound having thestructural wherein R is a substantially straight chain alkyl grouphaving 10 to l8 carbon atoms,

n is an integer 2 to 18;

R, R R and R are lower alkyls having 1 to 3 carbon atoms;

b is an integer 2 to l2;

X is selected from the group consisting of chlorine,

bromine and iodine;

wherein R is a substantially straight chain alkyl group having 10 to 18carbon atoms, i

Y is selected from the group consisting of 2 6 (5(m+ml m p and n is aninteger 2 to l8;

R, R R and R are lower alkyls having l to 3 carbon atoms;

b is an integer 2 to 12;

X is selected from the group consisting of chlorine,

bromine and iodine;

m is O to 3;

p is O to 3;

m+p is O to 3;

and

An is a compatible anion and a compatible inert pharmaceutical carriertherefor.

3. The method as set forth in claim 2 wherein R is n- C l-l and n is aninteger 3 to l0.

4. The method as set forth in claim 3 wherein Y is a -CH C H [5(m+p)] mp, m is zero and p is zero.

5. The method as set forth in claim 3 wherein is J CH C H [5-(r n-l-p)]m p, m is zero, p is l and X is chlorine.

.6. The antimicrobial composition as set forth in claim 1 wherein Y is(CH )l:l(;H C H R is n-C H and n is an integer 3 to 10. 7. Theantimicrobial composition as set forth in claim I wherein Y is (CH2)2N.(:H2C6H4Cl,

R is n-C H and n is an integer 3 to 10.

8. The method as set forth in claim 2 wherein R is n- CMH29' 9. Theantimicrobial composition as set forth in claim 8 wherein said compoundis [n-C H N*Me (CH N- *Me CH -C H -Cl]An wherein Me is methyl.

10. An antimicrobial composition comprising about 0.140% by weight of anunsymmetrical poly-onium quaternary ammonium compound having thestructural wherein R is a substantially straight chain alkyl grouphaving 10 to 18 carbon atoms,

Y is selected from the group consisting of n is an integer 2 to l8;

R. R R and R are lower alkyls having I to 3 carbon atoms:

b is an integer 2 to l2;

claim 10 wherein said pharmaceutical carrier is talc.

12. The antimicrobial composition as set forth in claim l2 wherein saidpharmaceutical carrier is clay.

13. The antimicrobial composition ase set forth in claim 10 wherein saidpharmaceutical carrier is diatomaceous earth.

14. The antimicrobial composition as set forth in claim 10 wherein saidpharmaceutical carrier is flour.

1. AN ANTIMICROBIAL COMPOSITION COMPRISING ABOUT 0.1-10% BY WEIGHT OF ANUNSYMMETRICAL POLY-ONIUM QUATERNARY AMMONIUM COMPOUND HAVING THESTRUCTURAL FORMULA:
 2. A method of inhibiting growth of microorganismscomprising applying to a surface infested with microorganisms anantimicrobial composition comprising about 0.1-10% by weight of anunsymmetrical poly-onium quaternary ammonium compound having thestructural formula:
 3. The method as set forth in claim 2 wherein R isn-C12H25, and n is an integer 3 to
 10. 4. The method as set forth inclaim 3 wherein Y is -CH2C6H (5-(m+p))RmXp, m is zero and p is zero. 5.The method as set forth in claim 3 wherein Y is -CH2C6H (5-(m+p))RmXp, mis zero, p is 1 and X is chlorine.
 6. The antimicrobial composition asset forth in claim 1 wherein Y is -(CH2)N -CH2C6H5, R is n-C12H25 and nis an integer 3 to
 10. 7. The antimicrobial composition as set forth inclaim 1 wherein Y is -(CH2)2N -CH2C6H4Cl, R is n-C12H25 and n is aninteger 3 to
 10. 8. The method as set forth in claim 2 wherein R isn-C14H29.
 9. The antimicrobial composition as set forth in claim 8wherein said compound is (n-C14H29N Me2(CH2)10N Me2CH2-C6H4p -Cl)Anwherein Me is methyl.
 10. An antimicrobial composition comprising about0.1-10% by weight of an unsymmetrical poly-onium quaternary ammoniumcompound having the structural formula:
 11. The antimicrobialcomposition as set forth in claim 10 wherein said pharmaceutical carrieris talc.
 12. The antimicrobial composition as set forth in claim 12wherein said pharmaceutical carrier is clay.
 13. The antimicrobialcomposition ase set forth in claim 10 wherein said pharmaceuticalcarrier is diatomaceous earth.
 14. The antimicrobial composition as setforth in claim 10 wherein said pharmaceutical carrier is flour.